Vybrant® FAM Poly Caspases Assay Kit, for flow cytometry

Catalog number: V35117

Molecular Probes™  Related applications: Cell Viability, Proliferation & Function | Cellular Imaging | Flow Cytometry | Target-Based ADME⁄Tox Assays

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Description

The Vybrant™ FAM Poly Caspases Assay Kit employs a novel approach to detect active caspases that is based on a fluorescent inhibitor of caspases (FLICA™) methodology, essentially an affinity label. The reagent associates a fluoromethyl ketone (FMK) moiety, which can react covalently with a cysteine, with a caspase-specific amine acid sequence. For poly caspases, this recognition sequence is valine-alanine-aspartic acid (VAD). A fluorescein group is attached as a reporter. The FLICA reagent is thought to interact with the enzymatic reactive center of an activated caspase via the recognition sequence, and then to attach covalently through the FMK moiety. The FLICA inhibitor is cell permeant and noncytotoxic. Unbound FLICA molecules diffuse out of the cell and are washed away; the remaining green-fluorescent signal is a direct measure of the amount of active caspase that was present at the time the inhibitor was added. This kit includes the FAM-VAD-FMK FLICA reagent, Hoechst 33342 stain, and propidium iodide stain, which allows the simultaneous evaluation of caspase activation, membrane permeability, and cell cycle by flow cytometry.

View a selection guide for all apoptosis assays for flow cytometry.
For Research Use Only. Not for use in diagnostic procedures.

Specifications

Form: Solution
Format: Tube(s)
Label or Dye: Propidium iodide, FMK, Hoechst 33342
Product Size: 25 assays
Detection Method: Fluorescent
For Use With (Equipment): Flow Cytometer
Excitation⁄Emission (nm): PI: 535⁄617, Hoechst 33342: 350⁄461, FAM-VAD-FMDK reagent: 488⁄530
Shipping Condition: Room Temperature

Contents & storage

Contains 1 vial of FAM-VAD-FMK caspase-8 reagent (FLICA reagent), 1 vial Hoechst 33342, 1 vial of propidium iodide, 1 vial of DMSO, 1 bottle of apoptosis fixative solution, and 1 bottle of apoptosis wash buffer (10X solution).

Store in refrigerator (2–8°C) and protect from light.

Documents

Manuals & protocols

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